Significance: In breast-preserving tumor surgery, the inspection of the excised tissue boundaries for tumor residue is too slow to provide feedback during the surgery. The discovery of positive margins requires a new surgery which is difficult and associated with low success. If the re-excision could be done immediately this is believed to improve the success rate considerably. Aim: Our aim is for a fast microscopic analysis that can be done directly on the excised tissue in or near the operating theatre. Approach: We demonstrate the combination of three nonlinear imaging techniques at selected wavelengths to delineate tumor boundaries. We use hyperspectral coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), and two-photon excited fluorescence (TPF) on excised patient tissue. Results: We show the discriminatory power of each of the signals and demonstrate a sensitivity of 0.87 and a specificity of 0.95 using four CARS wavelengths in combination with SHG and TPF. We verify that the information is independent of sample treatment. Conclusions: Nonlinear multispectral imaging can be used to accurately determine tumor boundaries. This demonstration using microscopy in the epi-direction directly on thick tissue slices brings this technology one step closer to clinical implementation.

中文翻译：

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用于肿瘤勾画的非线性多光谱成像。

意义：在保乳肿瘤手术中，对切除组织边界的肿瘤残留检查速度太慢，无法在手术过程中提供反馈。阳性切缘的发现需要一种新的手术，该手术难度大且成功率低。如果可以立即进行再切除，相信这将大大提高成功率。目标：我们的目标是进行快速的显微分析，可以直接在手术室内或附近的切除组织上进行。方法：我们展示了在选定波长下结合三种非线性成像技术来描绘肿瘤边界。我们在切除的患者组织上使用高光谱相干反斯托克斯拉曼散射 (CARS)、二次谐波生成 (SHG) 和双光子激发荧光 (TPF)。结果：我们展示了每个信号的辨别力，并使用四个 CARS 波长与 SHG 和 TPF 组合证明了 0.87 的灵敏度和 0.95 的特异性。我们验证信息与样本处理无关。结论：非线性多光谱成像可用于准确确定肿瘤边界。这种直接在厚组织切片上使用外向方向显微镜进行的演示使这项技术更接近临床实施。